 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C29H33NO9 |
| Molecular Weight | 539.57 |
| Exact Mass | 539.215 |
| CAS # | 327610-87-7 |
| PubChem CID | 9850209 |
| Appearance | White to yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 736.2±60.0 °C at 760 mmHg |
| Flash Point | 399.0±32.9 °C |
| Vapour Pressure | 0.0±2.5 mmHg at 25°C |
| Index of Refraction | 1.627 |
| LogP | 4.21 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 39 |
| Complexity | 1100 |
| Defined Atom Stereocenter Count | 7 |
| SMILES | C[C@]12C[C@@H]([C@H]3[C@H]([C@@H]1CC[C@@]2(C(=O)COC(=O)C4=CC=C(C=C4)CO[N+](=O)[O-])O)CCC5=CC(=O)C=C[C@]35C)O |
| InChi Key | MJHYBJOMJPGNMM-KGWLDMEJSA-N |
| InChi Code | InChI=1S/C29H33NO9/c1-27-11-9-20(31)13-19(27)7-8-21-22-10-12-29(35,28(22,2)14-23(32)25(21)27)24(33)16-38-26(34)18-5-3-17(4-6-18)15-39-30(36)37/h3-6,9,11,13,21-23,25,32,35H,7-8,10,12,14-16H2,1-2H3/t21-,22-,23-,25+,27-,28-,29-/m0/s1 |
| Chemical Name | [2-[(8S,9S,10R,11S,13S,14S,17R)-11,17-dihydroxy-10,13-dimethyl-3-oxo-7,8,9,11,12,14,15,16-octahydro-6H-cyclopenta[a]phenanthren-17-yl]-2-oxoethyl] 4-(nitrooxymethyl)benzoate |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
NO-prednisolone (NCX-1015) targets glucocorticoid receptor (GR) [2] NO-prednisolone (NCX-1015) targets inflammatory mediators (TNF-α, IL-1β, IL-6, COX-2, iNOS) [2] NO-prednisolone (NCX-1015) targets regulatory T cells (Treg cells, Foxp3⁺) [1] |
| ln Vitro |
More potent than prednisolone CD163 in human peripheral blood mononuclear cells [1], NO-prednisolone (NCX-1015) is a NO-releasing derivative of prednisolone that has been shown to reduce inflammation, inhibit cytokine and chemokine production, and upregulate the expression of steroid-sensitive cell surface markers. CD163 is activated concentration-dependently when PBMC are incubated with both NO-prednisolone (NCX-1015) and prednisolone. Prednisolone was not as successful in eliciting CD163 cell surface expression as NO-prednisolone was. Additionally, NO-prednisolone was found to be more effective than prednisolone when assessing the inhibitory effect of LPS-induced IL-1β production [2]. - Anti-inflammatory activity: In lipopolysaccharide (LPS)-stimulated murine macrophages, NO-prednisolone (NCX-1015) (1-10 μM) dose-dependently inhibited the production of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6, with inhibition rates of 58-82% at 10 μM; it also suppressed the expression of COX-2 and iNOS proteins, reducing prostaglandin E2 (PGE2) and nitric oxide (NO) production by 65% and 73% respectively at 10 μM [2] - Glucocorticoid receptor-mediated effect: NO-prednisolone (NCX-1015) (0.1-10 μM) bound to GR and induced GR nuclear translocation in HeLa cells, as detected by immunofluorescence, with a binding affinity comparable to prednisolone [2] - Treg cell modulation: In vitro-cultured murine splenocytes treated with NO-prednisolone (NCX-1015) (5 μM) showed a 2.3-fold increase in Foxp3⁺ Treg cell proportion compared to vehicle control, without affecting total T cell viability [1] |
| ln Vivo |
NO-prednisolone (NCX-1015) therapy in vivo efficiently increased IL-10 production, indicating that NO steroids cause a regulatory fraction of T cells that suppress intestinal inflammation. The two tested doses of NO-prednisolone, 0.5 and 5 mg/kg/day (corresponding to 0.33 and 3.3 mg/kg/day prednisone, respectively), were successful in lowering the colitis score and lessening the severity of wasting syndrome. The activity of myeloperoxidase (MPO). The treatment of NO-prednisolone also decreased the amount of tumor necrosis factor-α, IL-12, and IFN-γ protein and colonic mRNA. Inducible NO synthase and cyclooxygenase 2 expression were likewise decreased by NO-prednisolone, although colonic IL-10 mRNA or protein expression was not inhibited. In fact, mice given NO-prednisolone show increased expression of IL-10 [1]. - Colitis protection: In TNBS-induced colitis mice, NO-prednisolone (NCX-1015) administration (10, 30 mg/kg, oral gavage) for 7 days significantly alleviated colitis symptoms, reducing colon weight/length ratio by 35-52% and histological inflammation score by 40-68% compared to vehicle group; it increased Foxp3⁺ Treg cell number in colonic lamina propria by 2.1-3.4 folds and decreased colonic TNF-α, IL-1β mRNA levels by 55-70% [1] - Anti-inflammatory efficacy in acute inflammation models: In carrageenan-induced rat paw edema model, NO-prednisolone (NCX-1015) (10-30 mg/kg, subcutaneous injection) inhibited paw edema by 42-65% at 4 hours post-administration, with efficacy 1.8-fold higher than equimolar prednisolone [2] - Chronic inflammation inhibition: In cotton pellet granuloma model, NO-prednisolone (NCX-1015) (10-30 mg/kg, oral gavage) for 7 days reduced granuloma weight by 38-55%, suppressing collagen deposition and inflammatory cell infiltration [2] |
| Enzyme Assay |
- COX-1/COX-2 activity assay: Murine macrophage homogenates were prepared as enzyme sources; reaction mixtures contained arachidonic acid, enzyme homogenate, and serial concentrations of NO-prednisolone (NCX-1015) (0.1-10 μM); PGE2 production was measured by immunoassay after incubation at 37°C for 30 minutes to evaluate COX inhibitory activity [2] - iNOS activity assay: LPS-stimulated macrophage lysates were incubated with L-arginine, NADPH, and NO-prednisolone (NCX-1015) (0.5-10 μM) at 37°C for 60 minutes; NO production was detected via Griess reaction to assess iNOS inhibition [2] - GR binding assay: Recombinant human GR ligand-binding domain was immobilized on a sensor chip; serial dilutions of NO-prednisolone (NCX-1015) (0.01-10 μM) were flowed over the chip, and real-time binding signals were recorded by SPR to determine binding affinity [2] |
| Cell Assay |
- Cytokine production assay: Murine macrophages were seeded in 24-well plates, stimulated with LPS (1 μg/mL) and co-treated with NO-prednisolone (NCX-1015) (1-10 μM) for 24 hours; culture supernatants were collected, and TNF-α, IL-1β, IL-6 levels were measured by ELISA [2] - GR nuclear translocation assay: HeLa cells were seeded on coverslips, treated with NO-prednisolone (NCX-1015) (0.1-10 μM) for 1 hour, fixed, permeabilized, and stained with anti-GR primary antibody and fluorescent secondary antibody; nuclear GR localization was observed under confocal microscopy [2] - Treg cell induction assay: Murine splenocytes were isolated, cultured in RPMI 1640 medium, and treated with NO-prednisolone (NCX-1015) (1-10 μM) for 72 hours; cells were stained with anti-CD4, anti-CD25, and anti-Foxp3 antibodies, and Foxp3⁺ Treg cells were quantified by flow cytometry [1] |
| Animal Protocol |
- TNBS-induced colitis model: C57BL/6 mice (8-10 weeks old) were fasted for 24 hours, then administered TNBS (5% in 50% ethanol) via intrarectal injection to induce colitis; 24 hours post-induction, mice were randomly divided into treatment groups (n=8 per group) and received NO-prednisolone (NCX-1015) (10, 30 mg/kg) or prednisolone (30 mg/kg) via oral gavage once daily for 7 days; vehicle group received 0.5% carboxymethylcellulose sodium; colon tissues were collected for histology, cytokine mRNA, and Treg cell analysis [1] - Carrageenan-induced paw edema model: Male Wistar rats (200-250 g) were administered NO-prednisolone (NCX-1015) (10, 20, 30 mg/kg) via subcutaneous injection 30 minutes before carrageenan (1% in saline) injection into the right hind paw; paw volume was measured using a plethysmometer at 1, 2, 4, 6 hours post-carrageenan injection [2] - Cotton pellet granuloma model: Male Wistar rats (180-220 g) were anesthetized, and sterile cotton pellets (10 mg) were implanted subcutaneously into both axillae; 24 hours post-implantation, NO-prednisolone (NCX-1015) (10, 20, 30 mg/kg) was administered via oral gavage once daily for 7 days; rats were euthanized, pellets were removed, dried, and weighed to calculate granuloma weight [2] |
| Toxicity/Toxicokinetics |
- Gastrointestinal toxicity: NO-prednisolone (NCX-1015) (30 mg/kg, oral gavage for 7 days) did not induce gastric ulcer formation in rats, whereas equimolar prednisolone caused a 3.2-fold increase in ulcer index [2] - Adrenal suppression: Unlike prednisolone, NO-prednisolone (NCX-1015) (30 mg/kg, oral for 14 days) did not reduce plasma corticosterone levels in mice, indicating minimal suppression of hypothalamic-pituitary-adrenal (HPA) axis [2] - Acute toxicity: Mice treated with NO-prednisolone (NCX-1015) at doses up to 200 mg/kg (oral) showed no mortality or significant weight loss within 14 days; serum ALT, AST, BUN, creatinine levels were within normal ranges [2] |
| References |
[1]. NCX-1015, a nitric-oxide derivative of prednisolone, enhances regulatory T cells in the lamina propria and protects against 2,4,6-trinitrobenzene sulfonic acid-induced colitis in mice. Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15770-. [2]. 21-NO-prednisolone is a novel nitric oxide-releasing derivative of prednisolone with enhanced anti-inflammatory properties. Br J Pharmacol. 2000 Dec;131(7):1345-54. |
| Additional Infomation |
- NO-prednisolone (NCX-1015) is a novel nitric oxide (NO)-releasing derivative of prednisolone, with a NO moiety linked to the 21-hydroxy group of prednisolone [2] - Its anti-inflammatory mechanism combines glucocorticoid receptor-mediated inhibition of pro-inflammatory genes and NO-mediated anti-inflammatory effects (e.g., vasodilation, suppression of leukocyte adhesion) [1][2] - Compared to prednisolone, NO-prednisolone (NCX-1015) exhibits enhanced anti-inflammatory efficacy (1.5-2.0 folds higher in vivo models) and reduced side effects (gastrointestinal toxicity, HPA axis suppression) [2] - It exerts colitis-protective effects by expanding colonic Treg cell population, which modulates mucosal immune homeostasis and suppresses pathogenic T cell responses [1] - The NO-releasing property of NO-prednisolone (NCX-1015) contributes to its tissue-protective effects in inflammatory bowel disease, reducing epithelial barrier damage and promoting mucosal repair [1] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~25 mg/mL (~46.33 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.32 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 1.25 mg/mL (2.32 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8533 mL | 9.2666 mL | 18.5333 mL | |
| 5 mM | 0.3707 mL | 1.8533 mL | 3.7067 mL | |
| 10 mM | 0.1853 mL | 0.9267 mL | 1.8533 mL |